The research aimed to investigate the therapeutic effects and mechanisms of Opuntia dillenii Haw polysaccharide (OPS) on atherosclerosis of rats. First atherosclerotic rat models were established by high-fat and high-calcium diet. Thirty days later, the rats were treated with low dosage of OPS (0.2 g x kg(-1) x d(-1)) or high dosage of OPS (0.4 g x kg(-1) x d(-1)) by intraperitoneal injection for 60 days continuously. At the end of treatment, thoracic aorta rings were prepared and vasorelaxation of rat thoracic aorta in different experiment groups were determined by using 620M multi wire myograph system in vitro. Blood and livers of rats were collected. Then plasma levels of total cholesterol (TC), triglyceride (TG) and low density lipoprotein (LDL) of rats were separately determined using whole automatic biochemical analyzer; protein level of hepatic apolipoprotein B (ApoB) and that of hepatic diglyceride acyltransferase (Dgat1) were measured by Western Blot technique. Results showed that the ability of rat thoracic aorta to relax decreased markedly in the model group compared with that in the normal group, and significant differences existed in vasorelaxation ratios induced by different concentrations of carbamylcholine chloride (Carb) between these two groups (P < 0.01). After OPS treatment, the ability of rat thoracic aorta to relax improved markedly, the vasorelaxation ratios induced by Carb at 5 and 10 μmol x L(-1) were respectively 0.34 ± 0.08 and 0.62 ± 0.15 in the group treated with low dosage of OPS, while the ratios induced by Carb at 1 and 5 μmol x L(-1) were respectively 0.54 ± 0.08 and 0.98 ± 0.02 in the group treated with high dosage of OPS, which were all significantly different with those in the model group (P < 0.01). Plasma contents of TC, TG and LDL reduced significantly by the treatments both with low and high dosages of OPS compared with those in the model group (P < 0.01). Protein level of hepatic ApoB and that of hepatic Dgat1 decreased significantly after the treatment with high dosage of OPS compared with those in the model group (P < 0.01). These results indicate that OPS can markedly improve the vasorelaxation of thoracic aorta of atherosclerotic rats and has significant anti-atherosclerotic effect; inhibiting the expression of ApoB and Dgat1 and thus decreasing the amounts of TC, LDL and TG serving as one of the molecular mechanisms of its antiatherosclerosis effect.
Animals ; Aorta, Thoracic ; drug effects ; Atherosclerosis ; drug therapy ; Cholesterol ; blood ; Lipoproteins, LDL ; blood ; Opuntia ; chemistry ; Phytotherapy ; Rats ; Triglycerides ; blood

OBJECTIVETo establish a new culture method to induce the differentiation of embryonic pancreatic cells into mature endocrine cells.

METHODSMouse embryos at day 12.5 were used and embryonic pancreata were isolated. The isolated embryonic pancreata were cultured on the filter for 7 days, which floated in the dish containing medium. During culture, the expression of pancreas duodenum homeobox-1 (PDX-1), a pancreatic stem cell marker, was examined at day 1. The expression of neurogenin 3 (Ngn3), a pancreatic progenitor cell marker, was examined at day 3. The expressions of endocrine and exocrine markers, insulin, glucagon, and carboxypeptidase (CPA) were examined at day 7 by immunohistochemistry. The kinetics of pancreatic marker expression during culture was assayed by real-time PCR.

RESULTSMany pancreatic stem cells still existed in embryonic pancreata cultured for 1 day; meanwhile, these pancreatic stem cells proliferated in high rate. A large amount of pancreatic progenitor cells were found in embryonic pancreata cultured for 3 days.Pancreatic stem/progenitor cells differentiated into mature endocrine and exocrine cells in embryonic pancreata after having been cultured for 7 days. Furthermore, the expression pattern of pancreatic marker is consistent with that in vivo.

CONCLUSIONWe successfully established a new culture method, with which embryonic pancreatic cells can efficiently differentiate into mature endocrine cell.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Embryo, Mammalian ; Endocrine Cells ; cytology ; Female ; Homeodomain Proteins ; metabolism ; Male ; Mice ; Nerve Tissue Proteins ; metabolism ; Pancreas ; cytology ; Trans-Activators ; metabolism

Objective To further understand potential mechanism of miRNAs in bladder cancer .Mtehods Human microarray was used to analyze the expression of miRNAs in four pair human BC cancer tissues and adjacent normal tissues.Then RT-Qpcr was used to verified that the expression of two most up -regulated miRNAs and target genes for results of miRNA/Mrna microarray.Subsequently, it was deduced that miR-130b-3p could target at PTEN through a bioinformatics approach and dual-luciferase reporter assay.CCK8, EDU, flow cytometry, wound healing, Transwell and cytoskeleton assay were applied to prove that miR-130b could affect proliferation, apoptosis, migration and invasion of BC cells.The effects of PTEN regula-ted by miR-130b-3p on the key target protein expression of PI 3K/AKT and integrin β1/FAK signaling pathway were determined by Western blot.Resulst miR-130b-3p was significantly up-regulated and nega-tively correlated with PTEN expression in clinical bladder cancer specimens as compared with normal urothelial tissue.miR-130b-3p mimics could down-regulate PTEN expression, which leads to the activation of PI3K/AKT and integrinβ1/FAK signaling pathway related to cell proliferation , migration and invasion in bladder cancer EJ cells.When the cells were transfected with miR-130b-3p inhibitors, they could be sur- veyed with rearranging cytoskeleton.Conclusions miR-130b/PTEN may be used as a marker for bladder cancer.

AIMTo observe the role of melatonin receptor and GABAA receptor in sleeping time prolonged by melatonin in mice.

METHODSThe absence of the righting reflex was considered as the sleep onset and the duration of the loss of the righting reflex was recorded as the sleeping time. The effects of receptor agonist and antagonist on hypnotic activity of melatonin were studied in the paper.

RESULTSPrazosin hydrochloride, the blocker of melatonin 3 receptor, didn't affect the sleeping time prolonged by melatonin in mice. GABA, the endogenous agonist of GABA receptor, significantly potentiated the hypnotic activity of melatonin. When picrotoxin, the ligand of picrotoxin site on GABAA receptor, used together with melatonin, it significantly antagonized the sleeping time prolonged by melatonin, however, bicuculline, the specific antagonist of GABA binding site in GABAA receptor, didn't affect the hypnotic activity of melatonin in mice.

CONCLUSIONMelatonin does not exhibit its potentiation sleeping time in mice through melatonin 3 receptor. Hypnotic activity of melatonin may be mediated through picrotoxin site on GABAA receptor.

Animals ; Bicuculline ; pharmacology ; Male ; Melatonin ; physiology ; Mice ; Mice, Inbred Strains ; Picrotoxin ; pharmacology ; Prazosin ; pharmacology ; Receptors, GABA-A ; physiology ; Receptors, Melatonin ; physiology ; Sleep ; physiology

Aim:To investigate the effect of Hemangiopoietin (HAPO) on the hematopoiesis reconstitution in sub-lethally irradiated Balb/c mice.Methods: Balb/c mice were underwent total body irradiation at 700 cGy 137Cs ? radiation and were treated with HAPO or recombinant human granulocyte colony stimulating factor (rhG-CSF) after irradiation. The hematopoiesis reconstitution of mice were detected. Cells from bone marrow of Balb/c mice were cultured with HAPO or rhG-CSF for 24 hours or 72 hours before or after the cells were irradiated. The viability of cells were assessed and the ability of in vitro hematopoiesis reconstitution were also detected. Result: rhG-CSF and HAPO treated mice both showed increased survival rate and increased colony forming units. The peripheral WBC number increased greatly. The HAPO group was most quickest compared with rhG-CSF group and PBS control group. The number of bone marrow cells at day 14 of rhG-CSF group was higher than that in HAPO group, but the number of bone marrow cells at day 32 of rhG-CSF was lower than that in HAPO group. The number of bone marrow cells at day 42 of rhG-CSF was below normal. The number of bone marrow cells at day 42 of HAPO group was nearly normal. The number of CFU-GEMM in HAPO group was most compared with that in rhG-CSF group and PBS control group at day 7, 14 and 21 after radiation. The survival rate of cells after radiation in HAPO group was markedly higher than that in PBS control group, but the survival rate of cells after radiation in rhG-CSF group was no notable difference compared with that in PBS control group. In MTT assay, both HAPO and rhG-CSF incubation stimulated proliferation of bone marrow cell at 72 hours after radiation. Bone marrow cells formed Hematopoietic islands in HAPO group after radiation and were positive for sca-1 and CD31. CD31 positive endothelial cells increased around the Hematopoietic islands. There was no Hematopoietic islands formation, few CD31 positive endothelial cells and no sca-1 positive cells in PBS control group. Conclusion: HAPO can promote hematopoiesis reconstitution in sub-lethally irradiated Balb/c mice. It can increase the survival rate of mice and stimulate the proliferation of hematopoietic stem cells.

Aged ; Cardiac Catheters ; Combined Modality Therapy ; Coronary Restenosis ; therapy ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Paclitaxel ; administration & dosage ; therapeutic use ; Percutaneous Coronary Intervention ; instrumentation ; Stents ; Treatment Outcome ; Tubulin Modulators ; administration & dosage ; therapeutic use

OBJECTIVETo construct a method to culture pancreatic progenitor cells in hanging drop and on floating filter,and to examine if pancreatic progenitor cells can differentiate into mature endocrine cells with this method.

METHODSMurine embryos at day 12.5 were isolated and digested into single cells,which were then cultured in hanging drop for 24h and formed spheres.Spheres were cultured on the filter for 6 days,which floated in the dish containing medium.During culture,the expressions of pancreas duodenum homeobox-1(PDX-1)and neurogenin3(Ngn3)were determined.The expressions of endocrine and exocrine markers,insulin,glucagon,and carboxypeptidase(CPA)were determined on day 7 by immunohistochemistry.Insulin secretion of spheres stimulated by glucose was detected by ELISA.The changes of pancreatic marker expressions during culture were monitored by real-time polymerase chain reaction(PCR).

RESULTSOne day after the culture,there were still a large amount of PDX-1 positive cells in pancreatic spheres,and these cells proliferated.On day 3,high expression of Ngn3 was detected,and the Ngn3-positive cells did not proliferate.On day 7,The expressions of endocrine and exocrine markers in the differentiated pancreatic progenitor cells were detected,which were consistent with that in vivo.Insulin was secreted by spheres upon the stimulation of glucose.

CONCLUSIONIn hanging drop and on floating filter,pancreatic progenitor cells can differentiate into mature endocrine cells.

Animals ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Endocrine Cells ; cytology ; Homeodomain Proteins ; metabolism ; Insulin ; metabolism ; Mice ; Pancreas ; cytology ; Stem Cells ; cytology ; Trans-Activators ; metabolism

The aim of this study was to investigate the effects of interferon (IFN)-γ on biological characteristics and immunomodulatory property of human umbilical cord-derived mesenchymal stem cells (hUC-MSC). hUC-MSC were treated with IFN-γ 10 ng/ml (IFN-γ group) or without IFN-γ (control group). The phenotype of hUC-MSC was detected by flow cytometry. The proliferation status was detected by CCK-8 method, and its differentiation ability was assessed by oil red O and von Kossa staining. The production of PGE-2 was measured by ELISA, and the mRNA expression levels of COX-2, IDO-1 and IDO-2 in hUC-MSC were detected by real-time quantitative PCR. Furthermore, the proliferation of human peripheral blood mononuclear cells (hPBMNC) was evaluated after co-culture with hUC-MSC, IFN-γ pretreatment or not. The results showed that after IFN-γ stimulation, the expression of SSEA-4 on hUC-MSC decreased significantly [(8.15 ± 2.94) vs (16.42 ± 8.5), P < 0.05], and the expression of CD54 increased [(96.64 ± 3.29) vs (84.12 ± 10.73), P = 0.051]. The immunomodulatory property of hUC-MSC on the proliferation of hPBMNC was enhanced (P < 0.05). All the above mentioned effects were IFN-γ concentration-dependent. When hUC-MSC were stimulated by IFN-γ for 24 h, the production of PGE-2 secreted by hUC-MSC decreased significantly (P < 0.01). The mRNA expression level of COX-2 also decreased though the difference did not reach to statistically significant level. Compared with control group, IDO-1 expression level in IFN-γ group increased significantly (P < 0.01), and the mRNA expression level of IDO-2 remained unchanged. It is concluded that IFN-γ can influence the phenotype of hUC-MSC and enhance the immunomodulatory property of hUC-MSC.
Cells, Cultured ; Cyclooxygenase 2 ; metabolism ; Flow Cytometry ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; metabolism ; Interferon-gamma ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; drug effects ; immunology ; Umbilical Cord ; cytology

Objective To provide scientific evidences for improving children early integrated development,through analyzing the mental developmental status and characteristics of 322 cases of 2-year-old children in Nanjing.Methods Intelligence and motor development condition in 322 cases of 2-year-old children were assessed by using Bayley Scales of Infant Development test,and the assessed results were analyzed.Results 1.The incidences of the children whose mental development index(MDI)or psychomotor development index(PDI)were under 69 were 3.1% and 5.6%,respectively;2.The MDI mean score(114.34?19.65)was significantly higher than that of PDI(101.73?21.53)(t=9.71,P0.05).Conclusions The incidences of mental retardation in this study were consistent with the result reported by World Health Organization.There were differences between motor and intelligence development in children,as well as the intelligence development between male and female.Therefore,it should be implemented early childhood developmental screening in child health care.Parents should be given scientific guides about intelligence and motor development of children.

Hematopoietic stem cells (HSC) shift from fetal liver and spleen to bone marrow at neonatal stages and this movement may be due to inductive signals from different microenvironments. Mesenchymal stem cells (MSC) are the precursors of stromal cells in bone marrow microenvironments such as osteoblasts and endothelial cells. Some researchers speculated that fetal bone marrow before birth might be not perfectly suit HSC growth. However, it is still lack of direct evidence to prove this hypothesis. This study was aimed to compare the hematopoietic supportive capacity between human fetal and adult bone marrow MSC in vitro. Adult bone marrow MSC (ABM-MSC) were isolated from three healthy donors and fetal bone marrow MSC (FBM-MSC) were isolated from three fetuses between gestations of 19 to 20 weeks. After irradiation, MSC were co-cultured with CD34(+) cells isolated from umbilical cord blood in long-term culture-initiating cell (LTC-IC) assay. The colony number of colony forming cells (CFC) was counted and the phenotypic changes of co-cultured CD34(+) cells were analyzed by flow cytometry. Cytokine expressions in both kinds of MSC were detected by reverse transcription polymerase chain reaction (RT-PCR). The results showed that ABM-MSC had a stronger hematopoietic supportive capacity than FBM-MSC. Both of them enhanced the differentiation of CD34(+) cells into myeloid lineages. Cytokines were expressed differently in ABM-MSC and FBM-MSC. It is concluded that ABM-MSC possess more potential application in some treatments than FBM-MSC, especially in hematopoietic reconstitution.
Adult Stem Cells ; cytology ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Fetus ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology